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突变R107Q改变了mtSSB ssDNA的紧缩能力和结合动态

Martial Martucci¹, Amandine Moretton¹, Aleix Tarrés-Solé²

¹Université Clermont Auvergne, CNRS, Laboratoire de Physique de Clermont, F-63000 Clermont-Ferrand, France.

Nucleic acids research

|May 14, 2024

概括

R107Q突变损害了线粒体单链DNA结合蛋白 (mtSSB) 的功能,降低了它结合和紧单链DNA (ssDNA) 的能力. 这种分子缺陷与线粒体光学缩和视网膜缩有关.

科学领域:

线粒体生物学线粒体生物学
分子遗传学分子遗传学
生物化学生物化学

背景情况:

线粒体单链DNA结合蛋白 (mtSSB) 对于线粒体DNA (mtDNA) 复制至关重要.
mtSSB变种涉及到自体主导线粒体光学缩和视网膜缩.

研究的目的:

调查严重R107Q mtSSB变种背后的分子机制.
阐明R107Q突变对mtSSB与单链DNA (ssDNA) 相互作用的功能后果.

主要方法:

在体外分析mtSSB R107Q寡合状态.
单分子方法来评估ssDNA结合和紧缩.
实时竞争测定ssDNA结合的测定.
对ssDNA与mtSSB相互作用的分子建模.

主要成果:

在体外,mtSSBR107Q突变体在体外形成了稳定的四重体.
与野生类型 (WT) 的mtSSB相比,mtSSBR107Q表现出减少的ssDNA紧缩和增加的ssDNA解离.
在ssDNA结合竞争中,WT mtSSB在mtSSBR107Q上表现出明显的优势.
分子建模表明,R107Q突变破坏了静电相互作用,减少了ssDNA结合部位.

结论:

R107Q突变严重损害了mtSSB的ssDNA结合和紧缩能力.
这些功能性缺陷很可能是由于ssDNA包裹效率降低和蛋白质-ssDNA相互作用改变.
这些发现为mtSSB相关的线粒体疾病的发病提供了分子洞察力.

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